JP2005325031A - 1-cinnamylpiperidine derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compound useful as a preventing and treating agent of diseases to which a melanin-agglutinating hormone receptor inhibitor is effective, especially obesity, since it has a good ingestion-inhibiting activity based on the melanin-agglutinating hormone receptor-inhibiting effect. <P>SOLUTION: This 4-benzoylamino-1-cinnamylpiperidine derivative is provided by joining a nitrogen-containing saturated heterocyclic ring such as piperidine, etc., on the benzene ring of cinnamyl group through a linker. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a pharmaceutical, particularly a 1-cinnamylpiperidine derivative having a melanin-concentrating hormone receptor inhibitory action and useful as an anti-obesity drug, and a pharmaceutically acceptable salt thereof.

Obesity is defined as an increased state of adipose tissue weight, and is caused by a breakdown in the balance between energy intake and consumption. Modern lifestyle habits such as overeating and lack of exercise have led to a positive energy balance and are involved in the rapid increase in the obese population in countries around the world (see, for example, Non-Patent Document 1). In recent years, obesity has become a risk factor for lifestyle-related diseases including diabetes and hyperlipidemia, as well as mere body problems (see Non-Patent Document 1, for example).
Melanin-concentrating-Hormone (hereinafter referred to as MCH) is a cyclic peptide hormone isolated from the pituitary gland for the first time (see, for example, Non-Patent Document 2). Thereafter, the homologue was isolated also in rats, humans and the like (see, for example, Non-Patent Documents 3 and 4).
On the other hand, a gene called SLC-1 was obtained, and it was revealed that the gene encodes one of the MCH receptors (for example, see Non-Patent Document 5). SLC-1 is expressed in many regions of the brain including human and rat hippocampus, amygdaloid nucleus, thalamus, midbrain, pons, and hypothalamus (see, for example, Non-Patent Documents 6 and 7). In the hypothalamus, distribution is found in the arcuate nucleus, ventrolateral nucleus, dorsal medial nucleus, and solitary nucleus involved in feeding and energy balance control, suggesting that MCH may mediate energy homeostasis control (For example, see Non-Patent Document 8). In fact, the SLC-1 inhibitor suppressed the intake of MCH-induced food intake and favorite food, and suppressed weight gain due to a high fat diet (see, for example, Non-Patent Documents 9 and 10). Therefore, MCH receptor inhibitors, particularly SLC-1 inhibitors, are considered useful as anti-obesity agents.

(式中、Arは置換基を有していてもよく、縮合してもよい芳香環、X¹は置換基を有していてもよく、主鎖の原子数が1ないし5である2価の鎖状基、X⁴は結合手等、X²は非環式炭化水素基等、X³は結合手または置換基を有していてもよい2価の非環式炭化水素基、R²は塩基性置換基。詳細は当該公報参照。) So far, compounds having MCH receptor inhibitory action or SLC-1 inhibitory action have been reported. For example, Non-Patent Documents 9 and 10 disclose tetrahydronaphthalene derivatives and 4-phenylpiperidine derivatives, respectively. In addition, a compound represented by the following general formula has been reported (Patent Document 1).

(In the formula, Ar may have a substituent, may be condensed with an aromatic ring, X ¹ may have a substituent, and the main chain has a divalent valence of 1 to 5 atoms. X ⁴ is a bond, X ² is an acyclic hydrocarbon group, X ³ is a divalent acyclic hydrocarbon group which may have a bond or a substituent, R ² Is a basic substituent.

(式中、R₁〜R₈は同一若しくは互いに異なって、水素原子、ハロゲン、C₁-C₆アルキル又はC₁-C₆アルコキシ等、R₉〜R₁₉は独立して水素原子又はC₁-C₆アルキル、Wは窒素原子又はC-R_a、R_aは水素原子又は水酸基等、Xはハロゲン、シアノ又はニトロ等、Yは酸素原子、硫黄原子、-S(O)-又は-SO₂-、ZはC₁-C₆アルキル等。詳細は当該公報参照。) Further, it has been reported that a compound represented by the following general formula has an MCH receptor inhibitory action (Patent Document 2).

(Wherein R _{1 to} R ₈ are the same or different from each other, hydrogen atom, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, etc., R _{9 to} R ₁₉ are independently hydrogen atom or C ₁ -C ₆ alkyl, W is nitrogen atom or CR _a , R _a is hydrogen atom or hydroxyl group, X is halogen, cyano or nitro, Y is oxygen atom, sulfur atom, -S (O)-or -SO _2- Z represents C ₁ -C ₆ alkyl, etc. For details, refer to the publication.)

(式中、R¹はアルキル、アルケニル又はアリールアルケニル等[これらは有効な炭素原子を介してハロゲン、アルコキシ、アリールオキシ等から選択される1、2又は3個の基で置換されていてもよい]、R⁵は炭素数2以上のアルキル、アルケニル又はアリール等、R⁶は水素原子等。詳細は当該公報参照。)
当該公報には、該化合物のMCH受容体又はSLC-1の阻害活性に関して何らの記載もない。 On the other hand, compounds having a 4-benzoylamino-1-cinnamylpiperidine structure have been reported so far. For example, it has been reported that a compound represented by the following general formula has an inhibitory action on microsomal triglyceride transfer protein and is useful for the prevention or treatment of obesity (Patent Document 3).

(Wherein R ¹ is alkyl, alkenyl, arylalkenyl, etc. [these may be substituted with 1, 2, or 3 groups selected from halogen, alkoxy, aryloxy, etc. via an effective carbon atom] R ⁵ is an alkyl, alkenyl, aryl or the like having 2 or more carbon atoms, R ⁶ is a hydrogen atom, etc. For details, refer to the publication.)
There is no description in the publication regarding the inhibitory activity of the MCH receptor or SLC-1 of the compound.

(式中、Rはハロゲン原子又はヒドロキシ基等、R¹は水素原子又はアミノ基等、R²はニトロ基若しくはトリフルオロメチル基等、R⁸は水素原子等、R⁵〜R⁷はそれぞれが水素原子等、Wは低級アルケニル基等、R⁴中のR⁸〜R¹⁰は水素原子、若しくはハロゲン原子、又は低級アルコキシ等。詳細は当該公報参照。)
特許文献4〜8のいずれにおいても、該化合物のMCH受容体阻害作用に関する記載はなく、また、シンナミル基のベンゼン環上にリンカーを介して含窒素飽和ヘテロ環を有する化合物については、開示も示唆もない。 In addition, it has been reported that a compound described by the following general formula has dopamine antagonism and serotonergic activity and is useful for treatment of psychosis or mania (Patent Document 4). In Patent Documents 5 to 8, compounds having structures and actions similar to those of Patent Document 4 are described.

(In the formula, R is a halogen atom or a hydroxy group, R ¹ is a hydrogen atom or an amino group, R ² is a nitro group or a trifluoromethyl group, R ⁸ is a hydrogen atom, R ^{5 to} R ⁷ are each (A hydrogen atom, etc., W is a lower alkenyl group, etc., R ^{8 to} R ^{10 in} R ⁴ are a hydrogen atom, a halogen atom, lower alkoxy, etc. For details, refer to this publication.)
In any of Patent Documents 4 to 8, there is no description regarding the MCH receptor inhibitory action of the compound, and disclosure is also suggested for a compound having a nitrogen-containing saturated heterocycle via a linker on the benzene ring of the cinnamyl group. Nor.

Kopelman P.G. (2000) Nature 404, 635-643 Kawauchi, H. et al. (1983) Nature 305, 321-323 Nahon, J.L. et al. (1989) Endocrinology 125, 2056-2065 Presse, F. et al. (1990) Mol. Endocrinol. 4, 632-637 Saito, Y. et al. (1999) Nature 400, 265-269 Kolakowski L. F. et al. (1996) FEBS Lett. 398, 253-258 Lembo P. M. C. et al. (1999) Nature Cell Biol. 1, 267-271 Hervieu G. J. et al. (2000) Eur. J. Neurosci. 12, 1194-1216 Takekawa S. et al. (2002) Eur. J. Phramacol. 438, 129-135 Borowski B. et al. (2002) Nature Med. 8, 825-830 JP 2001-226269 A WO02 / 004433 pamphlet JP 07-165712 A JP 53-087365 A JP-A-53-090267 JP-A-53-087364 Irish Patent Invention No. 46270 Swiss patent application 628885

  An object of the present invention is to provide a novel anti-obesity drug having a good anti-feeding action based on an MCH receptor inhibitory action.

As a result of intensive studies on compounds having MCH receptor inhibitory activity, the present inventors have shown that nitrogen-containing saturated heterocycles such as piperidine are bonded to the benzene ring of the cinnamyl group via a linker, as shown in the following general formula. The characteristic 4-benzoylamino-1-cinnamylpiperidine derivative was confirmed to have a good anti-feeding activity based on the MCH receptor inhibitory action, and the pharmaceutical composition comprising this as an active ingredient was excellent. The present invention was completed by discovering that it can be an anti-obesity drug.
That is, the present invention relates to a novel 1-cinnamylpiperidine derivative represented by the following general formula (I) or a salt thereof.

(式中の記号は以下の意味を有する。
R^1a、R^1b及びR^1c:同一又は互いに異なって、H、低級アルキル、ハロゲン、ハロゲノ低級アルキル、-OH、-O-低級アルキル、-S-低級アルキル又は-NO₂、
R²:H又は低級アルキル、
X:-O-、-S-又は-NH-、
n:0、1、2、3又は4、
Het:単環式含窒素飽和ヘテロ環。以下同様。)

(The symbols in the formula have the following meanings.
R ^1a , R ^1b and R ^1c : the same or different from each other, H, lower alkyl, halogen, halogeno lower alkyl, -OH, -O-lower alkyl, -S-lower alkyl or -NO ₂ ,
R ² : H or lower alkyl,
X: -O-, -S- or -NH-,
n: 0, 1, 2, 3 or 4,
Het: Monocyclic nitrogen-containing saturated heterocycle. The same applies below. )

Since the compound of the present invention has a good antifeeding activity based on the MCH receptor inhibitory action, it is useful as an anti-obesity drug or a preventive or therapeutic drug for lifestyle-related diseases caused by obesity.
That is, as will be described later, the compound of the present invention exhibits an excellent MCH receptor inhibitory action and has a good inhibitory action on mouse fasting-induced feeding. In addition, this compound is expected as an effective anti-obesity drug having no QT prolonging action, which has been a problem in recent years in drug development.

Hereinafter, the present invention will be described in detail.
In the definition of the general formula in the present specification, the term “lower” means a straight or branched carbon chain having 1 to 6 carbon atoms (hereinafter abbreviated as C _1-6 ) unless otherwise specified. To do. Therefore, “lower alkyl” is C _1-6 alkyl, preferably linear alkyl of methyl, ethyl, propyl group, and branched alkyl such as isopropyl, butyl, isobutyl, tert-butyl group, etc. . Particularly preferred are the methyl, ethyl, propyl and isopropyl groups.
“Halogen” refers to F, Cl, Br and I. “Halogeno lower alkyl” preferably means C _1-6 alkyl substituted with one or more halogens, more preferably C _1-6 alkyl substituted with one or more F, still more preferably Are fluoromethyl, difluoromethyl, trifluoromethyl and trifluoroethyl groups.

  `` Monocyclic nitrogen-containing saturated heterocycle '' is a 4- to 8-membered saturated or partially unsaturated monocyclic group that contains one N atom and may further contain one heteroatom composed of N, S, and O. A ring heterocyclic group, preferably azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, azepanyl, diazepanyl, morpholinyl, thiomorpholinyl and tetrahydropyridyl groups, more preferably azetidinyl, pyrrolidinyl and piperidinyl groups.

(式中、R^1a〜R^1cは前記と同じ意味を有し、mは0又は1を、kは0又は1を示す。) In the compound represented by the general formula (I), preferred embodiments are as follows. R ^1a , R ^1b and R ^1c are preferably the same or different from each other, and H, methyl, ethyl, F, Cl, Br, -OMe or -S-Me, more preferably H, F, Cl or -OMe is there;
R ² is preferably H;
X is preferably -O-;
n is preferably 0;
Het is preferably azetidinyl, pyrrolidinyl or piperidinyl.
Particularly preferred compounds in the present invention are derivatives represented by the following (Ia) and salts thereof.

(Wherein R ^{1a to} R ^1c have the same meaning as described above, m represents 0 or 1, and k represents 0 or 1).

The compound of the present invention may have a tautomer based on rotation failure of the amide bond. The present invention also includes each of these isomers and mixtures.
In addition, the compound of the present invention may have an asymmetric carbon atom, and (R) -form and (S) -form optical isomers based on this may exist. The present invention includes all of these optical isomers and isolated ones. Further, the present invention includes compounds in which the compound of the present invention is labeled with a radioisotope.

Specific examples of the pharmaceutically acceptable salt of the compound of the present invention include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, and oxalic acid. Acid addition with organic acids such as acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, aspartic acid, glutamic acid Examples include salts.
Furthermore, the compound of the present invention includes pharmacologically acceptable prodrugs. A pharmacologically acceptable prodrug is a compound having a group that can be converted to NH ₂ , OH, CO ₂ H or the like of the present invention by solvolysis or under physiological conditions. Examples of groups that form prodrugs include those described in Prog. Med., 5, 2157-2161 (1985) and “Development of Pharmaceuticals” (Yodogawa Shoten, 1990), Volume 7, Molecular Design p163-198. .
Furthermore, the compound of the present invention and salts thereof include various hydrates, solvates and crystalline polymorphic substances.

(Production method)
The compound of the present invention and a salt thereof can be produced by applying various known synthesis methods utilizing characteristics based on the basic skeleton or the type of substituent. In this case, depending on the type of functional group, it is effective in terms of production technology to protect the functional group with an appropriate protective group at the raw material or intermediate stage, or to replace it with a group that can be easily converted to the functional group. There is a case. Examples of such a functional group include an amino group, a hydroxyl group, and a carboxyl group, and examples of protective groups thereof include, for example, Green (TW Greene) and Utz (PGM Wuts), "Protective Groups in Organic Synthesis (3rd edition, 1999) ”, which may be appropriately selected depending on the reaction conditions. In such a method, after carrying out the reaction by introducing the protecting group, the desired compound can be obtained by removing the protecting group as necessary or converting it to a desired group.
In addition, the prodrug of the compound (I) of the present invention is produced by introducing a specific group at the stage of the raw material or intermediate or reacting with the obtained compound (I) of the present invention in the same manner as the above protecting group. it can. The reaction can be carried out by applying a method known by those skilled in the art, such as ordinary esterification, amidation, dehydration and the like.
Although the typical manufacturing method of this invention compound or its intermediate is demonstrated below, it is not limited to these manufacturing methods.

(上記式中、X¹はOH又は脱離基[ハロゲン、1-ヒドロキシベンゾトリアゾール-1-イルオキシ、メトキシ、フェノキシ及びアジド基等]を表す。以下同様。)
本発明化合物(I)は、一般式(1)で表される化合物と一般式(2)で表される化合物とを、アミド化反応に供することにより得られる。
式(2)においてX¹がOHである化合物を用いる場合、本反応は、化合物(1)及び化合物(2)を等量或いは一方を過剰に用い、縮合試薬(ジシクロヘキシルカルボジイミド、1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド又は1,1'-カルボニルジイミダゾール(CDI)等)の存在下、反応に不活性な溶媒中、冷却下乃至加熱下で攪拌しながら行われる。上述の反応に不活性な溶媒としては、例えばジクロロメタン、1,2-ジクロロエタン(DCE)、クロロホルム、ベンゼン、トルエン、キシレン、エーテル、テトラヒドロフラン(THF)、ジオキサン、酢酸エチル(EtOAc)、エタノール、メタノール、2-プロパノール、アセトニトリル、N,N-ジメチルホルムアミド(DMF)、N,N-ジメチルアセトアミド、N-メチルピロリドン、ジメチルイミダゾリジノン、ジメチルスルホキシド(DMSO)、アセトン、メチルエチルケトン又は水等や、これらの均一系及び不均一系混合溶媒が挙げられるが、種々の反応条件に応じて適宜選択される。塩基(例えばトリエチルアミン(TEA)、ジイソプロピルエチルアミン及びピリジン等の有機塩基、又は炭酸カリウム及び炭酸水素ナトリウム等の無機塩基)の存在下、反応を行うことが好ましい場合がある。また、クロロギ酸エチルやクロロギ酸イソブチルを用いる混合酸無水物法等、通常アミド化に使用される縮合法を利用することもできる。
また、式(2)においてX¹が前記脱離基である反応性誘導体を用いる場合、前記同様の塩基の存在下或いは非存在下に、アミン化合物(1)と反応させる。このような反応性誘導体は対応するカルボン酸化合物から容易に導く事が可能であり、例えばX¹がハロゲンである反応性誘導体は、カルボン酸化合物をチオニルクロリド、オキザリルクロリド又はオキシ塩化リン等のハロゲン化試剤で処理すればよい。
以上のアミド化反応については、例えば、M.Bodanszky著「Peptide Chemistry」(1988年、p55-73)、泉屋信夫ら「ペプチド合成の基礎と実験」(1985年、p89-142)に詳細に記載されており、これらを参考に実施することができる。 (First production method)

(In the above formula, X ¹ represents OH or a leaving group [halogen, 1-hydroxybenzotriazol-1-yloxy, methoxy, phenoxy, azide group, etc.] and so on.)
The compound (I) of the present invention can be obtained by subjecting the compound represented by the general formula (1) and the compound represented by the general formula (2) to an amidation reaction.
In the case where a compound in which X ¹ is OH in the formula (2) is used, this reaction is carried out by using an equal amount or an excess of the compound (1) and the compound (2), and a condensation reagent (dicyclohexylcarbodiimide, 1-ethyl-3 In the presence of-(3-dimethylaminopropyl) carbodiimide or 1,1'-carbonyldiimidazole (CDI), etc.) in a solvent inert to the reaction while stirring under cooling to heating. Examples of the solvent inert to the above reaction include dichloromethane, 1,2-dichloroethane (DCE), chloroform, benzene, toluene, xylene, ether, tetrahydrofuran (THF), dioxane, ethyl acetate (EtOAc), ethanol, methanol, 2-propanol, acetonitrile, N, N-dimethylformamide (DMF), N, N-dimethylacetamide, N-methylpyrrolidone, dimethylimidazolidinone, dimethylsulfoxide (DMSO), acetone, methyl ethyl ketone, water, etc. System and heterogeneous mixed solvents can be mentioned, and they are appropriately selected according to various reaction conditions. It may be preferred to carry out the reaction in the presence of a base (eg, an organic base such as triethylamine (TEA), diisopropylethylamine and pyridine, or an inorganic base such as potassium carbonate and sodium bicarbonate). Further, a condensation method usually used for amidation, such as a mixed acid anhydride method using ethyl chloroformate or isobutyl chloroformate, can also be used.
In addition, when a reactive derivative in which X ¹ is the leaving group in the formula (2) is used, it is reacted with the amine compound (1) in the presence or absence of the same base as described above. Such a reactive derivative can be easily derived from the corresponding carboxylic acid compound. For example, a reactive derivative in which X ¹ is halogen is a carboxylic acid compound such as thionyl chloride, oxalyl chloride or phosphorus oxychloride. What is necessary is just to process with a halogenated reagent.
The above amidation reaction is described in detail, for example, in “Peptide Chemistry” by M. Bodanszky (1988, p55-73), Nobuo Izumiya et al. “Basics and Experiments of Peptide Synthesis” (1985, p89-142). It can be implemented with reference to these.

本発明化合物(I)は、一般式(3)で表されるアミン化合物と一般式(4)で表されるホルミル化合物とを、還元的アミノ化反応に供することにより得られる。
本反応では、まず、化合物(1)及び(2)を等量或いは一方を過剰に用い、第1製造法に記載の反応に不活性な溶媒中、冷却下乃至加熱下で攪拌し脱水縮合を行う。次いで、反応系に水素化シアノホウ素ナトリウム、水素化トリアセトキシホウ素ナトリウム、水素化ホウ素ナトリウム又はギ酸等の還元剤を加え、冷却下乃至加熱下攪拌して還元反応を行う。酸(例えばチタニウムテトライソプロポキシド等のルイス酸、酢酸又はp-トルエンスルホン酸等)の存在下に反応させるのが、反応を円滑に進行させる上で有利な場合がある。上述の還元については水素雰囲気下或いはギ酸アンモニウム等の水素供与剤存在下、パラジウム(Pd)、又は白金(Pt)等の金属触媒を用いて前記の反応に不活性な溶媒中接触水素還元することによっても行われる。
還元剤の添加時期は化合物(3)及び化合物(4)の混合直後に入れても、また、時間をおいて入れても良い。
以上の還元的アミノ化反応に関しては、例えば、日本化学会編「実験化学講座(丸善)」(第4版、20巻、1992年、p300)に詳細に記載されており、これを参考に実施することができる。 (Second production method)

The compound (I) of the present invention can be obtained by subjecting an amine compound represented by the general formula (3) and a formyl compound represented by the general formula (4) to a reductive amination reaction.
In this reaction, first, an equal amount of compound (1) and (2) or an excess of one of them is used, and the mixture is stirred under cooling or heating in a solvent inert to the reaction described in the first production method to perform dehydration condensation. Do. Next, a reducing agent such as sodium cyanoborohydride, sodium triacetoxyborohydride, sodium borohydride or formic acid is added to the reaction system, and the reaction is stirred under cooling to heating to carry out a reduction reaction. The reaction in the presence of an acid (for example, a Lewis acid such as titanium tetraisopropoxide, acetic acid, p-toluenesulfonic acid, etc.) may be advantageous in order to facilitate the reaction. For the above reduction, catalytic hydrogen reduction in a solvent inert to the above reaction using a metal catalyst such as palladium (Pd) or platinum (Pt) in a hydrogen atmosphere or in the presence of a hydrogen donor such as ammonium formate. It is also done by.
The reducing agent may be added immediately after mixing of the compound (3) and the compound (4), or may be added after some time.
The above reductive amination reaction is described in detail in, for example, “Chemical Experiment Course (Maruzen)” edited by The Chemical Society of Japan (4th edition, Volume 20, 1992, p300). can do.

(上記式中、X²は脱離基[ハロゲン、メタンスルホニルオキシ又はp-トルエンスルホニルオキシ等]、X³はWittig試薬又はHorner-Emmons試薬に用いられる基[ホスホニウム塩又は亜リン酸ジエステル等]を表す。以下同様。)
本反応は、等量から過剰量の塩基(例えばTEA及びジイソプロピルエチルアミン等の有機塩基又は炭酸カリウム、炭酸ナトリウム及び炭酸セシウム等の無機塩基)存在下、化合物(3)、化合物(5)及び化合物(6)を第1製造法に記載の反応に不活性な溶媒中、等量またはいずれかを過剰量用いて、冷却下乃至加熱下攪拌しながら行われる。添加剤(ヨウ化テトラ-n-ブチルアンモニウム又はヨウ化カリウム等)の存在下に反応させるのが、反応を円滑に進行させる上で有利な場合がある。
先に化合物(3)と化合物(5)とを反応させ、一旦中間体を単離した後、化合物(6)との反応を行っても良い。また、先に化合物(5)と化合物(6)とを反応させ、一旦中間体を単離した後、化合物(3)との反応を行っても良い。 (Third production method)
The compound (I) of the present invention comprises an amine compound represented by the general formula (3), a Wittig reagent or Horner-Emmons reagent represented by the general formula (5), and an aldehyde compound represented by the general formula (6). And can be produced by combining alkylation and olefination reactions.

(In the above formula, X ² is a leaving group [halogen, methanesulfonyloxy, p-toluenesulfonyloxy and the like], X ³ is a group used for Wittig reagent or Horner-Emmons reagent [phosphonium salt or phosphite diester, etc.] (The same applies hereinafter.)
This reaction is carried out in the presence of an equivalent amount to an excess amount of a base (for example, an organic base such as TEA and diisopropylethylamine or an inorganic base such as potassium carbonate, sodium carbonate, and cesium carbonate) (3), (5) and ( 6) is carried out while stirring under cooling to heating, using an equal amount or an excess of either in an inert solvent for the reaction described in the first production method. The reaction in the presence of an additive (such as tetra-n-butylammonium iodide or potassium iodide) may be advantageous for allowing the reaction to proceed smoothly.
The compound (3) and the compound (5) may be reacted first to isolate the intermediate, and then the reaction with the compound (6) may be performed. Alternatively, the compound (5) and the compound (6) may be reacted first to isolate the intermediate, and then reacted with the compound (3).

(上記式中、P¹及びP²はアミノ基の保護基を表す。) (Production method of raw material compound)
The starting compound (1) or (3) is obtained from the corresponding starting compound by the reductive amination reaction of the second production method or the amidation reaction of the first production method, or the alkylation and olefination of the third production method. It can be produced by the following reaction route using the same method as the reaction.

(In the above formula, P ¹ and P ² represent amino protecting groups.)

(上記式中。X⁴はフルオロ基又はクロロ基等の脱離基を表す。以下同様。) The starting compound (6) can be produced according to the following reaction route. Here, the substitution reaction is carried out with stirring under cooling or heating, using an equivalent amount or an excess amount in one in a solvent inert to the reaction. Bases (e.g. inorganic bases such as potassium carbonate, sodium carbonate and cesium carbonate, organic bases such as TEA and diisopropylethylamine, metal alkoxides such as potassium tert-butoxide and sodium tert-butoxide, or sodium hydride and lithium hydride) In some cases, the reaction in the presence is advantageous in order for the reaction to proceed smoothly. The reduction reaction is described in detail in, for example, “Chemical Experiment Course (Maruzen)” edited by the Chemical Society of Japan (4th edition, Volume 21, 1991, p89), and can be carried out with reference to these. .

(In the above formula, X ⁴ represents a leaving group such as a fluoro group or a chloro group; the same shall apply hereinafter.)

(上記式中、P³はカルボキシル基の保護基を表す。) The starting compound (4) can be produced according to the following reaction route. Here, the condensation reaction is the method described in the Chemical Society of Japan `` Experimental Chemistry Course (Maruzen) '' (4th edition, volume 19, 1992, p57). (4th edition, volume 21, 1991, p83), and the oxidation reaction is "Chemical Experiment Course (Maruzen)" edited by the Chemical Society of Japan (4th edition, volume 21, 1991, p2). Each of the methods described in (1) can be carried out with reference.
In the reduction reaction, a compound obtained by removing the protecting group of compound (15) to form a carboxyl group, directly reducing this, or converting the carboxyl group into a reactive derivative by the method described in the first production method It can also be carried out by a method of reducing after guiding.

(In the above formula, P ³ represents a protecting group for a carboxyl group.)

The compound of the present invention or the raw material compound thus produced is isolated as it is or as a salt thereof. The salt of the said compound can be manufactured by attaching | subjecting a normal salt-forming reaction to this invention compound which is a free base. The compound of the present invention or a salt thereof is isolated and purified as a hydrate, a solvate or a crystalline polymorphic substance. Isolation / purification is performed by applying ordinary chemical operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, and various chromatography.
Various isomers can be isolated by conventional methods utilizing the difference in physicochemical properties between isomers. For example, optical isomers can be obtained by a method of fractional recrystallization after introducing a racemate into a diastereomeric salt with an optically active organic acid (such as tartaric acid), or a column chromatography using a chiral filler, respectively. It can be separated and purified. The optically active compound can also be produced by using an appropriate optically active compound as a raw material.

A pharmaceutical composition containing one or more compounds of the present invention or a salt thereof as an active ingredient is prepared using carriers, excipients, and other additives that are usually used for formulation.
Administration is oral by tablet, pill, capsule, granule, powder, liquid, etc., or parenteral administration by injection such as intravenous injection, intramuscular injection, suppository, transdermal agent, nasal agent or inhalant. Either form may be sufficient. The dose is appropriately determined according to the individual case, taking into account the symptoms, age of the subject, sex, etc., but is usually about 0.001 mg / kg to 100 mg / kg per day for oral administration. This is administered once or divided into 2 to 4 times. In addition, when intravenously administered depending on symptoms, it is usually administered once or several times a day in the range of 0.0001 mg / kg to 10 mg / kg per adult. In the case of inhalation, it is usually administered once or multiple times per day in the range of 0.0001 mg / kg to 1 mg / kg per adult.
As the solid composition for oral administration according to the present invention, tablets, powders, granules and the like are used. In such solid compositions, one or more active substances are present in at least one inert excipient such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, metasilicate. Mixed with magnesium aluminate acid. The composition may contain an inert additive, for example, a lubricant such as magnesium stearate, a disintegrant such as sodium carboxymethyl starch, and a solubilizing agent according to a conventional method. If necessary, tablets or pills may be coated with a sugar coating or a gastric or enteric coating agent.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, etc., and include commonly used inert solvents such as purified water, ethanol, etc. . In addition to the inert solvent, the composition may contain adjuvants such as solubilizers, wetting agents, and suspending agents, sweeteners, corrigents, fragrances, and preservatives.
Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of the aqueous solvent include distilled water for injection and physiological saline. Examples of the non-aqueous solvent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, alcohols such as ethanol, polysorbate 80 (trade name), and the like. Such a composition may further contain isotonic agents, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, and solubilizing agents. These are sterilized by, for example, filtration through a bacteria-retaining filter, blending with a bactericide or irradiation. These can also be used by producing a sterile solid composition and dissolving and suspending it in sterile water or a sterile solvent for injection before use.

Hereinafter, the present invention will be described in more detail based on examples. The compounds of the present invention are not limited to the compounds described in the following examples. Moreover, the manufacturing method of a raw material compound is shown in a reference example. In addition, the compounds shown in Table 7 below can be easily produced by substantially the same methods as those described in the following Examples and the above production methods, or by applying some variations obvious to those skilled in the art from those methods. Can do.
The symbols in Examples, Reference Examples and Tables below have the following meanings.
Rf .: Reference number; Ex .: Example number; Structure: Structural formula; Salt: Addition salt; Data: Physicochemical properties (FA: FAB-MS (M + 1); EI: EI-MS (M +) ESI: ESI-MS (M + 1); LC: LC-MS (M + 1); N: NMR (DMSO-d ₆ , TMS internal standard) characteristic peak δ ppm); NO .: Compound number; Me : Methyl; Et: ethyl; iPr: isopropyl; Boc: tert-butoxycarbonyl; fum: fumarate; 2HCl: dihydrochloride;

Reference example 1
(±) -4-Hydroxyazepane-1-carboxylic acid Add 3.36 g of tert-butoxypotassium to a solution of 5.16 g of tert-butyl tert-butyl and stir at 60 ° C. for 1 hour, and then add 2.42 g of 4-fluorobenzonitrile. And stirred at 60 ° C. for 2 hours. After the reaction solution was cooled to room temperature, the solvent was distilled off under reduced pressure, and the residue was diluted with EtOAc. After washing with water and saturated brine, the organic layer was dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: hexane = 1: 4 → 1: 3) to obtain 4.36 g of tert-butyl (±) -4- (4-cyanophenoxy) azepane-1-carboxylate as colorless oil. Obtained. FA: 317
Reference example 2
(±) -4- (4-Cyanophenoxy) azepane-1-carboxylic acid Hydrogenated in a solution of 4.30 g of tert-butyl tert-butyl in 80 ml of toluene while stirring at -78 ° C so that the temperature of the reaction solution does not exceed -60 ° C. 41 ml of diisobutylaluminum chloride (1.0 mol / l toluene solution) was added dropwise. After stirring at −78 ° C. for 2 hours, methanol and saturated aqueous sodium potassium tartrate were added dropwise at the same temperature, and the temperature was slowly raised to room temperature. The insoluble material was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (EtOAc: hexane = 1: 4) to give (±) -4- (4- 4.06 g of tert-butyl formylphenoxy) azepane-1-carboxylate was obtained. FA: 320
Reference Example 3
To a suspension of 1.4 g of sodium hydride in 50 ml of THF was added 6.3 ml of ethyl diethylphosphonoacetate under ice cooling and stirred for 30 minutes, and then 8.1 g of tert-butyl 4- (4-formylphenoxy) piperidine-1-carboxylate, A 30 ml THF solution was added dropwise. After gradually warming to room temperature and stirring overnight, a saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with EtOAc. The organic layer was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (EtOAc: hexane = 1: 6 → 1: 4) to give colorless oil 4- {4 There was obtained 8.23 g of tert-butyl-[(1E) -3-ethoxy-3-oxoprop-1-en-1-yl] phenoxy} piperidine-1-carboxylate. FA: 376
Reference example 4
4- {4-[(1E) -3-Ethoxy-3-oxoprop-1-en-1-yl] phenoxy} piperidine-1-carboxylic acid Hydrogenation of 8.2 g of tert-butyl tert-butyl in 44 ml of THF at -78 ° C with stirring 51 ml of a 1.0M diisobutylaluminum toluene solution was gradually added dropwise within a range where the internal temperature did not exceed -60 ° C, and the mixture was stirred for 2 hours. A saturated aqueous ammonium chloride solution was added to the reaction mixture, the temperature was raised to room temperature, EtOAc was added, and the mixture was washed with 5% potassium hydrogen sulfate solution, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and 7.2 g of tert-butyl 4- {4-[(1E) -3-hydroxyprop-1-en-1-yl] phenoxy} piperidine-1-carboxylate as a pale yellow powder was removed. Obtained. FA: 334

Reference Example 5
4- {4-[(1E) -3-Hydroxyprop-1-en-1-yl] phenoxy} piperidine-1-carboxylic acid Manganese (IV) oxide in a 50 ml dichloromethane solution of tert-butyl 2.00 g under room temperature stirring 5.00 g was added and stirred overnight. After filtration through celite, the solvent was distilled off under reduced pressure to give 4- {4-[(1E) -3-oxoprop-1-en-1-yl] phenoxy} piperidine-1-carboxylic acid as a pale yellow powder. 2.00 g of tert-butyl acid was obtained. FA: 332
Reference Example 6
4- {4-[(1E) -3-oxoprop-1-en-1-yl] phenoxy} piperidine-1-carboxylate tert-butyl piperidin-4-ylcarbamate 6.64 g and 3.44 ml of acetic acid were added and stirred at room temperature for 1.5 hours, and then 12.73 g of sodium triacetoxyborohydride was added and stirred at room temperature for 65 hours. The reaction solution was diluted with 200 ml of EtOAc and washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine. The organic layer was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was washed with diisopropyl ether to give a colorless solid 4- [4-((1E) -3- {4-[(tert- 7.83 g of tert-butyl butoxycarbonyl) amino] piperidin-1-yl} prop-1-en-1-yl) phenoxy] piperidine-1-carboxylate were obtained. FA: 516
Reference Example 7
4- [4-((1E) -3- {4-[(tert-butoxycarbonyl) amino] piperidin-1-yl} prop-1-en-1-yl) phenoxy] piperidine-1-carboxylic acid tert- To a solution of butyl 7.82 g in EtOAc (20 ml) was added 4M hydrochloric acid-EtOAc (60 ml) under ice cooling, and the mixture was stirred for 3 hours while gradually warming to room temperature. After distilling off the solvent under reduced pressure, the residue was washed with EtOAc. The obtained compound was dissolved in 200 ml of chloroform and washed with 1M aqueous sodium hydroxide solution. The organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to give 1-{(2E) -3- [4- (piperidin-4-yloxy) phenyl] prop-2-ene as a pale yellow oil. -1-yl} piperidin-4-amine 7.83 g was obtained. FA: 316
Reference Example 8
To a solution of 1.61 g of 1-{(2E) -3- [4- (piperidin-4-yloxy) phenyl] prop-2-en-1-yl} piperidin-4-amine in 20 ml of benzene is added 0.54 g of benzaldehyde. It stirred for 5 hours, removing the water to produce | generate by azeotropy under heating reflux. After cooling the reaction solution to room temperature, 1.11 g of di-tert-butyl dicarbonate was added and stirred at room temperature for 16 hours. Next, after adding 6.6 ml of 1M potassium hydrogen sulfate aqueous solution and stirring at room temperature for 2 hours, 0.88 g of concentrated aqueous ammonia was added and stirred at room temperature for 1 hour. After extracting three times with 30 ml of chloroform, the organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol: concentrated aqueous ammonia = 90: 10: 1), and colorless solid 4- {4-[(1E) -3- (4-aminopiperidin-1-yl) 1.57 g of tert-butyl prop-1-en-1-yl] phenoxy} piperidine-1-carboxylate was obtained. FA: 416

Reference Example 9
0.62 g of potassium carbonate and 0.44 ml of 2-iodopropane were added to a solution of 0.51 g of methyl 4-fluoro-3-hydroxybenzoate in 15 ml of acetonitrile, stirred at room temperature for 24 hours, and then stirred at 60 ° C. for 11 hours. The reaction mixture was cooled to room temperature, diluted with water, and extracted with EtOAc. The organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (EtOAc: hexane = 1: 10) to obtain 0.66 g of methyl 4-fluoro-3-isopropoxybenzoate as colorless oil. EI: 212
Reference Example 10
To a solution of 0.59 g of methyl 4-fluoro-3-isopropoxybenzoate in 14 ml of methanol was added a 7 ml solution of 1M aqueous sodium hydroxide solution, and the mixture was stirred at 60 ° C. for 14 hours. After cooling the reaction solution to room temperature, the solvent was distilled off under reduced pressure, and the residue was acidified with 1M aqueous hydrochloric acid solution. The precipitate was collected by filtration and washed with water to obtain 0.55 g of colorless solid 4-fluoro-3-isopropoxybenzoic acid. FA: 197 (M-1)
Reference Example 11
To a solution of 0.74 g of 3-chloro-5-methoxybenzoic acid in 10 ml of DMF, 0.99 g of N-ethyl-N'-3-dimethylaminopropylcarbodiimide hydrochloride and 0.70 g of 1-hydroxybenzotriazole were added. The mixture was stirred for 1 hour, 0.79 g of tert-butyl 4-aminopiperidine-1-carboxylate was added, and the mixture was stirred at room temperature for 18 hours. After the solvent was distilled off under reduced pressure, the residue was washed with EtOAc to obtain 1.24 g of tert-butyl 4-[(3-chloro-5-methoxybenzoyl) amino] piperidine-1-carboxylate as a colorless solid. ESI: 369
Reference Example 12
4-[(3-Chloro-5-methoxybenzoyl) amino] piperidine-1-carboxylic acid To a solution of tert-butyl 1.22 g in EtOAc (3 ml) was added ice-cooled 4 M hydrochloric acid-EtOAc (9 ml). Stir for hours. After distilling off the solvent under reduced pressure, the residue was washed with ethyl acetate to obtain colorless solid of 3-chloro-5-methoxy-N-piperidin-4-ylbenzamide hydrochloride. This compound was dissolved in 1M aqueous sodium hydroxide solution, extracted with chloroform, and the organic layer was dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was washed with a mixed solvent of EtOAc-hexane to obtain 0.88 g of colorless solid 3-chloro-5-methoxy-N-piperidin-4-ylbenzamide. ESI: 269
Reference examples 13-35
The compound of Reference Example 13 was prepared in the same manner as in Reference Example 1, the compound of Reference Example 14 was prepared in the same manner as in Reference Example 2, the compounds of Reference Examples 15 to 18 were prepared in the same manner as in Reference Example 3, and Reference Example 4 The compounds of Reference Examples 19 to 22 were prepared in the same manner as described above, the compounds of Reference Examples 23 to 26 were prepared in the same manner as in Reference Example 5, the compounds of Reference Example 27 were prepared in the same manner as in Reference Example 10, and the references 11 and 11 were used. The compounds of Reference Examples 28 to 31 were produced in the same manner, and the compounds of Reference Examples 32 to 35 were produced in the same manner as in Reference Example 12. The structural formulas and physicochemical properties of the compounds of Reference Examples 13 to 35 are shown in Table 2 below.

Example 1
To a solution of 0.26 g of 3-chloro-4-fluoro-N-piperidin-4-ylbenzamide in 10 ml of 2-propanol, 0.45 g of (2-chloroethyl) triphenylphosphonium bromide, 0.70 g of potassium carbonate, tetra-n-iodide 0.07 g of butylammonium and 0.28 g of tert-butyl 3- (4-formylphenoxy) azetidine-1-carboxylate were added, and the mixture was stirred for 16 hours with heating under reflux. After cooling to room temperature, the solvent was distilled off under reduced pressure. The residue was diluted with EtOAc and washed with water and saturated brine. The organic layer was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol: concentrated aqueous ammonia = 98: 2: 0.2) to give an oily 3- [4-((1E) -3- {4-[(3-Chloro-4-fluorobenzoyl) amino] piperidin-1-yl} prop-1-en-1-yl) phenoxy] azetidine-1-carboxylic acid 0.11 g of tert-butyl was obtained.
To a solution of 0.106 g of the obtained compound in 2 ml of EtOAc, 4 ml of 4M hydrochloric acid-EtOAc was added under ice cooling, and the mixture was stirred for 2 hours while slowly warming to room temperature. After evaporating the solvent under reduced pressure, the residue was washed with a mixed solvent of ethyl acetate-methanol, and colorless solid N- (1-{(2E) -3- [4- (azetidin-3-yloxy) phenyl] Propa-2-en-1-yl} piperidin-4-yl) -3-chloro-4-fluorobenzamide hydrochloride 0.042 g was obtained.
Example 2
3-Chloro-4-fluorobenzoic acid 87 mg and 4- {4-[(1E) -3- (4-aminopiperidin-1-yl) prop-1-en-1-yl] phenoxy} piperidine-1-carvone To a solution of tert-butyl acid 207 mg in DMF 10 ml, N-ethyl-N′-3-dimethylaminopropylcarbodiimide hydrochloride 143 mg and 1-hydroxybenzotriazole 114 mg were added, and the mixture was stirred at room temperature for 19 hours. After evaporating the solvent under reduced pressure, the residue was diluted with EtOAc and washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine. The organic layer was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to give a pale yellow amorphous powder of 4- [4-((1E) -3- {4-[(3-chloro-4-fluoro Benzyl) amino] piperidin-1-yl} prop-1-en-1-yl) phenoxy] piperidine-1-carboxylate 254 mg was obtained.
Boc was removed from the obtained compound 185 mg in the same manner as in Example 1, and amorphous powder 3-chloro-4-fluoro-N- (1-{(2E) -3- [4- (piperidine -4-yloxy) phenyl] prop-2-en-1-yl} piperidin-4-yl) benzamide hydrochloride 176 mg was obtained.

Example 3
3-Chloro-4-fluoro-N-piperidin-4-ylbenzamide 0.13g and (3S) -3- {4-[(1E) -3-oxoprop-1-en-1-yl] phenoxy} pyrrolidine-1 -0.09 ml of acetic acid was added to a solution of 0.16 g of tert-butyl carboxylate in 5 ml of dichloromethane and stirred at room temperature for 1.5 hours, and then 0.32 g of sodium triacetoxyborohydride was added and stirred at room temperature for 14 hours. After evaporating the solvent under reduced pressure, the residue was diluted with EtOAc and washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine. The organic layer was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol: concentrated aqueous ammonia = 98: 2: 0.2) to give (3S) -3- [4-((1E) -3- {4- 0.24 g of tert-butyl [(3-chloro-4-fluorobenzoyl) amino] piperidin-1-yl} prop-1-en-1-yl) phenoxy] pyrrolidine-1-carboxylate was obtained.
Boc was removed from 0.23 g of the obtained compound in the same manner as in Example 1, and colorless solid 3-chloro-4-fluoro-N- [1-((2E) -3- {4-[(3S ) -Pyrrolidin-3-yloxy] phenyl} prop-2-en-1-yl) piperidin-4-yl] benzamide 0.158 g of hydrochloride was obtained.
Example 4
0.28 g of acetic acid was added to 0.124 g of 3-chloro-4-fluoro-N-piperidin-4-ylbenzamide and (2E) -3- [4- (3-pyrrolidin-1-ylpropoxy) phenyl] acrylaldehyde in 5 ml of THF. After adding ml and stirring at room temperature for 30 minutes, 0.31 g of sodium triacetoxyborohydride was added and stirred at room temperature for 5 hours. The reaction mixture was made alkaline by adding an aqueous sodium hydrogen carbonate solution, extracted with EtOAc, and the organic layer was dried over anhydrous sodium sulfate. After distilling off the solvent under reduced pressure, the residue was purified by silica gel column chromatography (chloroform: methanol: concentrated aqueous ammonia = 10: 1: 0.1) to give 3-chloro-4-fluoro-N- (1 0.23 g of-((2E) -3- [4- (3-pyrrolidin-1-ylpropoxy) phenyl] prop-2-en-1-yl) piperidin-4-yl) benzamide was obtained. 0.046 g of fumaric acid was added to an ethanol solution of 0.21 g of this compound and stirred at room temperature for 1 hour. After distilling off the solvent under reduced pressure, the residue was washed with EtOAc, and colorless solid 3-chloro-4-fluoro-N- (1-{(2E) -3- [4- (3-pyrrolidine-1- 0.161 g of ylpropoxy) phenyl] prop-2-en-1-yl} piperidin-4-yl) benzamide fumarate was obtained.
Examples 5-36
The compound of Example 5 was prepared in the same manner as in Example 1, the compound of 6 to 25 was prepared in the same manner as in Example 2, the compound of Examples 26 to 35 was prepared in the same manner as in Example 3, and the compounds of Examples The compound of Example 36 was produced in the same manner as in 4. The structural formulas and physicochemical properties of each Example compound are shown in Tables 3 to 6 below.

Example 37
(i) Isolation of mouse SLC-1 gene, cloning into expression vector and preparation of SLC-1 stable expression cell Full-length cDNA encoding mouse SLC-1 (SEQ ID NO: 3) was obtained by PCR. For amplification of the gene encoding SLC-1, 5'-ggaaagcttgccgccatggatytgcaagcctcgttgc-3 '(SEQ ID NO: 1) as a forward primer and 5'-ggactcgagtcaggtgcctttgctttctgtc-3' (SEQ ID NO: 1) as a reverse primer using cDNA derived from mouse brain as a template 2) was used. For PCR, PfuTurbo DNA Polymerase (Stratagene) was used, and a cycle of 94 ° C. (30 seconds) / 60 ° C. (30 seconds) / 74 ° C. (1 minute) was repeated 35 times. As a result, a DNA fragment having an open reading frame of 1062 bases (SEQ ID NO: 3) was amplified. This fragment was inserted into the HindIII / XhoI site of pcDNA3.1 / Zeo (+) (Invitrogen) to construct a mouse SLC-1 expression vector.
TypeI Collagen 10cm culture dish coated with (Asahi Techno Glass Co., Ltd.) to 293 after the cells were seeded in 24-hour culture at 1x10 ⁶ cells, pcDNA3.1 / Hygro of 0.8μg (+) (Invitrogen Corp.) and 7.2μg pSRE of -Luc (Clontech) was transfected with FuGENE6 (Boeringer Mannheim). 40 hours after gene transfer, the cells were seeded again in a 10 cm culture dish, selected with 100 μg / ml hygromycin (Boehringer Mannheim), and the cells that survived and formed colonies were collected to obtain SRE-lucierase gene stably transfected cells . Furthermore, the mouse SLC-1 expression vector was similarly introduced into this cell using FuGENE6 (Boeringer Mannheim), selected with 40 μg / ml of zeocin (Invitrogen), and the mouse SLC-1 stable expression cell of the present invention Got.

(ii) Measurement of mouse SLC-1 antagonist activity using luciferase reporter assay of test substance The mouse SLC-1 stably expressing cells produced were seeded at ⁹ × 10 ³ per well in a 96-well plate and cultured at 37 ° C. for 24 hours. . A test substance was added, and after incubation at 37 ° C. for 10 minutes, MCH was added to a final concentration of 50 nM. At this time, a test substance non-added group and an MCH non-stimulated group were prepared and used to calculate the SLC-1 activation inhibition rate of the test substance. After reaction at 37 ° C for 4 hours, the medium was discarded, and a luciferin substrate solution (3.8 mM Tricine-containing solution containing cell lysate (12.5 mM Tris HCl (pH 7.8), 1 mM DTT, 5% glycerol, 0.5% Triton-X100)) was obtained. , 0.3 mM MgCO ₃ , 0.5 mM MgSO ₄ , 20 μM EDTA, 24 mM DTT, 0.015% CoenzymeA, 40 μM ATP, 0.01% luciferin) was added and dissolved and reacted. Subsequently, the activity of luciferase was measured using a luminometer (ML3000; Dynatech Laboratories). SLC-1 activation inhibition rate (%) = (Luciferase activity when MCH is added to the test substance-luciferase activity when nothing is added) / (Luciferase activity when only MCH is added-nothing is added) The IC ₅₀ value of the test substance was calculated from the inhibition rate as luciferase activity x100 when not.
The compound of the present invention showed an excellent melanin-concentrating hormone receptor inhibitory action. The results for representative example compounds are shown in the table below.

実施例38
絶食誘発摂食に対する作用
(i)動物
雄性ddyマウス(7-9週令,日本SLC株式会社)を使用した。動物は標準飼育条件下(明期 7:30 - 20:30, 室温23 ± 2 ℃, 湿度55 ± 10 % )で飼育し,給餌および飲水は自由に行わせた。全ての実験は明期に実施した。
(ii)絶食誘発摂食実験
入荷1週間後に個別飼育ケージに移し、1週間慣化した後に実験を実施した。16時間絶食したマウスに化合物もしくはvehicleを皮下もしくは経口投与し、投与1時間後に餌を与え、給餌から1時間後までの摂食量を測定した。
(iii)化合物溶液調整
化合物は4-12% DMSO(関東化学株式会社)および4-12%クレモホール(ナカライテスク株式会社)を含む生理食塩水もしくは蒸留水に溶解、もしくは0.5% メチルセルロース(純正化学株式会社)を含む蒸留水に懸濁した。
(iv)データ解析
結果は平均値±S.E.M.で表示した。統計ツールにはSAS (ver. 6.11)を使用し, vehicle群との比較はDunnett's testで行った。
本発明化合物は良好な摂食抑制作用を示した。

Example 38
Effects on fasting-induced feeding
(i) Animals Male ddy mice (7-9 weeks old, Japan SLC Co., Ltd.) were used. Animals were reared under standard rearing conditions (light period 7:30-20:30, room temperature 23 ± 2 ℃, humidity 55 ± 10%), and they were allowed to freely feed and drink water. All experiments were conducted during the light period.
(ii) Fasting-induced feeding experiment One week after arrival, the animal was transferred to an individual breeding cage and the experiment was conducted after acclimatization for one week. A compound or vehicle was subcutaneously or orally administered to mice fasted for 16 hours, and food was given 1 hour after administration, and the amount of food intake from 1 hour after feeding was measured.
(iii) Preparation of compound solution The compound is dissolved in physiological saline or distilled water containing 4-12% DMSO (Kanto Chemical Co., Ltd.) and 4-12% Cremophor (Nacalai Tesque Co., Ltd.), or 0.5% methylcellulose (Pure Chemical Co., Ltd.) Suspended in distilled water.
(iv) Data analysis The results were expressed as mean ± SEM. SAS (ver. 6.11) was used as a statistical tool, and Dunnett's test was used for comparison with the vehicle group.
The compound of the present invention showed a good feeding inhibitory action.

  The description of “Artificial Sequence” is described in the numerical heading <223> of the following sequence listing. Specifically, each base sequence represented by the sequences of SEQ ID NOs: 1 and 2 in the sequence listing is an artificially synthesized primer sequence.

Claims (1)

A 1-cinnamylpiperidine derivative represented by the general formula (I) or a salt thereof.

(The symbols in the formula have the following meanings.
R ^1a , R ^1b and R ^1c : the same or different from each other, H, lower alkyl, halogen, halogeno lower alkyl, -OH, -O-lower alkyl, -S-lower alkyl or -NO ₂ ,
R ² : H or lower alkyl,
X: -O-, -S- or -NH-,
n: 0, 1, 2, 3 or 4,
Het: Monocyclic nitrogen-containing saturated heterocycle. )